Selective Acoustic Trapping, Translating, Rotating, and Orienting of Organism From Heterogeneous Mixture.

Selective contactless manipulation of organisms with intrinsic mobility from heterogeneous mixture is essential for biomedical engineering and microbiology. Acoustic manipulation, compared to its optical, magnetic, and electrostatic counterparts, provides superior bio-compatibility and additive-free properties. In this study, we present an acoustic manipulation system capable of selectively trapping, translating, rotating, and orienting individual organisms from in-Petri dish organism mixture using a phased transducer array and microscope, by dynamically steering the acoustic field. Specifically, using brine shrimp and zebrafish populations as example, the to-be-manipulated organisms with different sizes or morphologies can be manually designated by the user in microscopic image and interactively localized. Thereafter, the selected organisms can be automatically trapped from the heterogeneous mixture using a multiple focal point-based acoustic field steering method. Finally, the trapped organisms can be translated, rotated, and oriented in regard to the user's distinct manipulation objectives in instant response. In different tasks, closed-loop positioning and real-time motion planning control are performed, highlighting the innovation in terms of automation and accuracy of our manipulation technique. The results demonstrate that our acoustic manipulation system and acoustic field steering method enable selective, stable, precision, real-time, and in-Petri dish manipulation of organisms from heterogeneous mixture.

OXPHOS capacity is diminished and the phosphorylation system inhibited during diapause in an extremophile, embryos of Artemia franciscana.

Diapause exhibited by embryos of Artemia franciscana is accompanied by severe arrest of respiration. A large fraction of this depression is attributable to downregulation of trehalose catabolism that ultimately restricts fuel to mitochondria. This study now extends knowledge on the mechanism by revealing metabolic depression is heightened by inhibitions within mitochondria. Compared with that in embryo lysates during post-diapause, oxidative phosphorylation (OXPHOS) capacity P is depressed during diapause when either NADH-linked substrates (pyruvate and malate) for electron transfer (electron transfer capacity, E) through respiratory Complex I or the Complex II substrate succinate are used. When pyruvate, malate and succinate were combined, respiratory inhibition by the phosphorylation system in diapause lysates was discovered as judged by P/E flux control ratios (two-way ANOVA; F1,24=38.78; P<0.0001). Inhibition was eliminated as the diapause extract was diluted (significant interaction term; F2,24=9.866; P=0.0007), consistent with the presence of a diffusible inhibitor. One candidate is long-chain acyl-CoA esters known to inhibit the adenine nucleotide translocator. Addition of oleoyl-CoA to post-diapause lysates markedly decreased the P/E ratio to 0.40±0.07 (mean±s.d.; P=0.002) compared with 0.79±0.11 without oleoyl-CoA. Oleoyl-CoA inhibits the phosphorylation system and may be responsible for the depressed P/E in lysates from diapause embryos. With isolated mitochondria, depression of P/E by oleoyl-CoA was fully reversed by addition of l-carnitine (control versus recovery with l-carnitine, P=0.338), which facilitates oleoyl-CoA transport into the matrix and elimination by ß-oxidation. In conclusion, severe metabolic arrest during diapause promoted by restricting glycolytic carbon to mitochondria is reinforced by depression of OXPHOS capacity and the phosphorylation system.

Prepared microplastics interaction with Artemia salina under low pH conditions representing ocean acidification; a simulated environmental exposure.

Ocean acidification and microplastic pollution are two of the major ecological concerns. The distribution of large quantities of plastic debris and microplastics all across the oceans emphasises the need to determine the influence of microplastics in ocean acidification and to evaluate its concomitant toxicological effects on aquatic life forms. Studies on the combined impact of both the stressors are very limited, but much needed in the current scenario. Where most of the present-day research use purchased microplastics of defined size and morphology (microspheres, fibres, rods, etc.), the present study employs prepared "true to life microplastics" that resemble the environmental microplastic pollutants in morphology and size heterogeneity. The present study focusses on evaluating the fate and impact of oceanic microplastics on the physiology and development of Artemia salina (Brine shrimp), one among the most ecologically significant zooplankton species. Natural sea water was acidified by controlled perturbation of carbon dioxide using a valve system. The hatching rate of A. salina cysts receded significantly (p < 0.05) upon singular exposures to microplastics and low pH (7.80), whereas combined effect was insignificant. The reactive oxygen species (ROS) elevated as a result of individual exposures to microplastics and low pH. However, only in 0.5 mg mL-1 PE treatments at pH 7.80, an additive impact was reported for ROS activity (p < 0.05). The SOD activities increased significantly but it can be attributed as the individual responses towards exposure to both the stressors. A significant additive impact was not observed for SOD activity (p > 0.05). But during the development, significant morphological anomalies were observed. Changes in the appendages of nauplii and juveniles as a result of combined exposure to microplastics and low pH treatments are significant findings. Our observations suggest that coupled exposure to microplastics and low pH could induce significant oxidative stress in the marine zooplanktons and also adversely affect their normal development. Findings from the current study emphasise the need for further research to understand the coupled toxicological impacts of ocean acidification and predominant pollutants such as microplastics to other marine animals as well.

Reversible intracellular acidification and depletion of NTPs provide a potential physiological origin for centuries of dormancy in an Antarctic freshwater copepod.

A great diversity of crustacean zooplankton found in inland and coastal waters produce embryos that settle into bottom sediments to form an egg bank. Embryos from these banks can remain dormant for centuries, creating a reservoir of genetic diversity. A large body of literature describes the ecological and evolutionary importance of zooplankton egg banks. However, literature on the physiological traits behind dormancy in crustacean zooplankton are limited. Most data on the physiology of dormancy comes from research on one species of anostracan, the brine shrimp, Artemia franciscana. Anoxia-induced dormancy in this species is facilitated by a profound and reversible acidification of the intracellular space. This acidification is accompanied by a reversible depletion of adenosine triphosphate (ATP). The present study demonstrates that acidification of the intracellular space also occurs in concert with a depletion of nucleoside triphosphates (NTPs) in the Antarctic copepod, Boeckella poppei. Like A. franciscana, the depletion of NTPs and acidification are rapidly reversed during aerobic recovery in B. poppei. These data provide the first comparative evidence that extreme dormancy under anoxia in crustacean zooplankton is associated with intracellular acidification and an ability to recover from the depletion of ATP.

Impact of test chamber design on spontaneous behavioral responses of model crustacean zooplankton Artemia franciscana.

The use of small aquatic model organisms to investigate the behavioral effects of chemical exposure is becoming an integral component of aquatic ecotoxicology research and neuroactive drug discovery. Despite the increasing use of invertebrates for behavioral phenotyping in toxicological studies and chemical risk assessments, little is known regarding the potential for environmental factors-such as geometry, size, opacity and depth of test chambers-to modulate common behavioral responses. In this work, we demonstrate that test chamber geometry, size, opacity and depth can affect spontaneous, unstimulated behavioral responses of euryhaline crustacean Artemia franciscana first instar larval stages. We found that in the absence of any obvious directional cues, A. franciscana exhibited a strong innate wall preference behavior. Using different test chamber sizes and geometries, we found both increased wall preference and lowered overall distance traveled by the test shrimp in a smaller chamber with sharper-angled vertices. It was also determined through quantifiable changes in the chambers' color that the A. franciscana early larval stages can perceive, differentiate and react to differences in color or perhaps rather to light transmittance of the test chambers. The interaction between innate edge preference and positive phototaxis could be consistently altered with a novel photic stimulus system. We also observed a strong initial preference for depth in A. franciscana first instar larval stages, which diminished through the acclimatization. We postulate that the impact of test chamber designs on neurobehavioral baseline responses warrants further investigation, in particular considering the increased interest in behavioral eco-neurotoxicology applications.

Lethal, behavioral, growth and developmental toxicities of alkyl-PAHs and non-alkyl PAHs to early-life stage of brine shrimp, Artemia parthenogenetica.

Alkyl-PAHs are the predominant form of PAHs in crude oils which are supposed to demonstrate different toxicities compared to non-alkyl PAHs. Little information is available about the toxicity of alkyl-PAHs on marine Artemia. This study addressed and compared the lethal, behavioral, growth and developmental toxicities of three alkyl-PAHs, namely 3-methyl phenanthrene (3-mPhe), retene (Ret) and 2-methyl anthracene (2-mAnt), to their non-alkyl forms, phenanthrene (Phe) and anthracene (Ant) using Artemia parthenogenetica (nauplii, <24 h) as test organism following a 48 h and a 7 d of exposure, respectively. Benzo-a-pyrene (Bap) was selected as a reference toxicant for the comparison with the above alkyl-PAHs and non-alkyl PAHs. Results showed that for all tested endpoints, A. parthenogenetica nauplii had the highest sensitivity to Bap while Ant had no significant effect on nauplii survival or development within given concentrations. Considering the aqueous freely dissolved PAH concentrations, the 48 h-LC50 (survival), 48 h-EC50 (immobility) and 7 d-LC10 (survival) of Bap were calculated as 0.321, 0.285 and 0.027 µg/L, respectively, which were twofold to fivefold lower than those of Phe, 3-mPhe, Ret, Ant and 2-mAnt. A higher acute toxicity of alkyl-PAHs (3-mPhe and 2-mAnt) than their non-alkyl forms (Phe and Ant) was observed. Not limited to Phe, the common non-polar narcotic mode of action was also observed for Bap, 3-mPhe, Ret and 2-mAnt, which was evident by the inhibited mobility of nauplii. The decreased body lengths were found for all PAH treatments compared to the solvent control, whereas instar retardations were only found in nauplii exposed to Bap, Phe and Ret. Our findings emphasized the sensitivity differences of A. parthenogenetica nauplii to selected alkyl PAHs and non-alkyl PAHs and confirmed the application of lethal, behavioral and growth indicators in the toxicity evaluation of selected PAHs other than Ant. However, the distinct toxicities of these PAHs suggested other toxic modes of action may play more important roles apart from narcotic mode of action and need to be elucidated in future studies. In addition, a strong correlation between the body length and the instar of A. parthenogenetica nauplii was observed for each PAH exposure, suggesting that body length can be representative for both growth and developmental indicators during biological monitoring of PAH pollution in marine environment.

Effects of commercial formulations of glyphosate on marine crustaceans and implications for risk assessment under temperature changes.

Glyphosate-based formulations are the most commonly used herbicides worldwide with the risk of potential contamination of aquatic bodies. The present study assessed the response of four marine crustaceans to three different brands of herbicides Roundup®Platinum, Efesto® and Taifun® MK CL.T, under two selected temperatures of 20 °C and 30 °C. The harpacticoid copepod Tigriopus fulvus, the anostracan Artemia franciscana, the amphipod Corophium insidiosum and the isopod Sphaeroma serratum were chosen as testing organisms. Effects of herbicides and temperatures were assessed by estimating lethal concentrations. The results showed that the high temperature rises the toxicity of glyphosate with an increase of mortality of all the tested species. This is an important aspect for future risk assessments of pesticides under global climate change scenarios. Efesto® resulted the most toxic brand, showing C. insidiosum the most sensitive with 96 h-LC50 values of 3.25 mg/L acid equivalent (a.e.) at 30 °C and 7.94 mg/L a.e. at 20 °C followed by T. fulvus while A. franciscana and S. serratum were the less sensitive. This study provides important information for assessing the toxic effects of three different brands of glyphosate-based herbicides on non-target marine organisms suggesting that they should be carefully managed to minimize any negative impact on marine organisms.

Short exposure of Artemia salina to group-12 metals: Comparing hatchability, mortality, lipid peroxidation, and swimming speed.

The hatchability, mortality rate, lipid peroxide levels, and swimming speed of Artemia salina have been compared based on short exposures of ZnCl2, CdCl2, and HgCl2 in artificial seawater. The hatching tests were carried out for 12, 24, 36, and 48 h at 28 °C. Mortality rate and lipid peroxide (LPO) levels were determined after 24 h of exposure at 28 °C, in the dark, and on living larvae using the FOX method. The swimming speed was determined after 24 h using a microcomputer coupled to a digital camera, with simultaneous treatment of the recorded images every 25 s, at 25 °C, under red-light irradiation. Results showed that Zn caused a gradual inhibition of the hatching for concentrations <900 µmol L-1; however, Cd and Hg displayed almost complete inhibition for concentrations ≤100 µmol L-1. Also, the heavy metals caused a dose-dependent increase of mortality (LD50) in the following order: Zn = 3290 µmol L-1 < Cd = 2206 µmol L-1 < Hg = 15.6 µmol L-1. Furthermore, significant LPO levels were found for Cd (1500-2000 µmol L-1, p < 0.001) and Hg (5-20 µmol L-1, p < 0.001). Finally, the swimming speed values increased significantly, for Zn ≈ 2.5 mm s-1 (1500 µmol L-1, p < 0.001), Cd ≈ 3.5 mm s-1 (2000 µmol L-1, p < 0.05), and Hg ≈ 4.0 mm s-1 (15 µmol L-1, p < 0.05), after 24 h exposure. There is a clear dose-dependent toxicity, indicating that Zn, Cd and Hg can induce significant changes in hatchability, mortality, and ethological and biochemical parameters.

Reproductive performance in successive generations of the brine shrimp Artemia salina (Crustacea: Anostraca) from the Sebkha of Sidi El Hani (Tunisia).

Artemia salina cysts collected from the Sebkha of Sidi El Hani hatched and cultivated in the laboratory were investigated. Nauplii were acclimated to laboratory conditions until reproductively mature (Generation 1; G1). Reproductive performance in terms of total cysts and nauplii offspring, days between brood production, and cyst and nauplii numbers was evaluated. Nauplii produced by specimens in the G1 were defined as those of G2 with there also being third (G3) and fourth (G4) generations. The aims were to evaluate Artemia salina in aquaculture conditions with the long-term aim of developing a system for "artemiculture". There was a larger number of cysts or nauplii per brood between G1 (60.7 ±â€¯10.9 nauplii or cysts/female) and subsequent generations, notably G2 (122 nauplii or cysts/female, P <  0.05). The mean number of cysts and nauplii per female increased from the first and last brood. Number of cysts produced per female increased markedly (P <  0.05) from G1 (54.5 ± 14.2) to G3 (128.9 ± 39.2). Percentage of females producing nauplii increased from G1 (20 %) to G4 (100 %). There was, therefore, an increase in percentage of ovoviviparous offspring (nauplii) per female (P <  0.05) from G1 (7%) to G4 (71 %). There were no differences, however, between mean numbers of broods per female, and numbers of days between broods. The results indicate there are variations in reproduction from oviparity to ovoviviparity as consecutive generations of Artemia salina are cultivated. In summary, there was a rapid adaptation of Artemia salina of Sidi El Hani to laboratory conditions.

Effects of feeding on the physiological performance of the stony coral Pocillopora acuta.

Reef-building corals rely on both heterotrophy and endosymbiotic dinoflagellate autotrophy to meet their metabolic needs. Those looking to culture these organisms for scientific or industrial purposes must therefore consider both feeding regimes and the light environment. Herein the effects of three photosynthetically active radiation (PAR) levels were assessed in fed and unfed specimens of the model coral Pocillopora acuta that were cultured in a recirculating aquaculture system (RAS). Half of the corals were fed Artemia sp. brine shrimp in a separate feeding tank to prevent biofouling, and fragments were exposed to PAR levels of 105, 157, or 250 µmol quanta m-2 s-1 over a 12-h period each day. All cultured corals survived the 140-day treatment, and the physiological response variables assessed-buoyant weight, specific growth rate, linear extension, color, and Fv/Fm-were significantly influenced by feeding, and, to a lesser extent, light. Specifically, fed corals grew faster and larger, and presented darker pigmentation; corals fed at the highest light levels grew at the fastest rate (6 cm year-1 or 175 mg g-1 week-1). Given the high physiological performance observed, we advocate the active feeding of brine shrimp in RAS by those looking to cultivate P. acuta, and likely other corals, over long-term timescales.

Short-term cold stress and heat shock proteins in the crustacean Artemia franciscana.

In their role as molecular chaperones, heat shock proteins (Hsps) mediate protein folding thereby mitigating cellular damage caused by physiological and environmental stress. Nauplii of the crustacean Artemia franciscana respond to heat shock by producing Hsps; however, the effects of cold shock on Hsp levels in A. franciscana have not been investigated previously. The effect of cold shock at 1 °C followed by recovery at 27 °C on the amounts of ArHsp90, Hsp70, ArHsp40, and ArHsp40-2 mRNA and their respective proteins in A. franciscana nauplii was examined by quantitative PCR (qPCR) and immunoprobing of western blots. The same Hsp mRNAs and proteins were also quantified during incubation of nauplii at their optimal growth temperature of 27 °C. qPCR analyses indicated that the abundance of ArHsp90, Hsp70, and ArHsp40 mRNA remained relatively constant during both cold shock and recovery and was not significantly different compared with levels at optimal temperature. Western blotting revealed that ArHsp90, ArHsp40, and ArHsp40-2 were generally below baseline, but at detectable levels during the 6 h of cold shock, and persisted in early recovery stages before declining. Hsp70 was the only protein that remained constant in quantity throughout cold shock and recovery. By contrast, all Hsps declined rapidly during 6 h when nauplii were incubated continuously at 27 °C optimal temperature. Generally, the amounts of ArHsp90, ArHsp40, and ArHsp40-2 were higher during cold shock/recovery than those during continuous incubation at 27 °C. Our data support the conclusion that low temperature preserves Hsp levels, making them available to assist in protein repair and recovery after cold shock.

Assessment of the use of cooked Araucaria angustifolia seed coats extract as food for brine shrimp.

The use of vegetable waste and its screening for potential cytotoxicity is of utmost importance to ensure its safe use in the feed industry for fish and other animals. We evaluated aqueous and ethanolic extracts of cooked Araucaria angustifolia seed coats. The Stiasny index for the aqueous and ethanolic extracts was 2.87% ± 0.03% and 60.53% ± 4.79%, respectively. Condensed tannins were 11-fold higher in the ethanolic extract than the aqueous extract. The flavonoid and polyphenol contents were 1.7- and 1.8-fold higher in the ethanolic extract than in the aqueous extract, respectively. The 36 h EC50 for brine shrimp hatchability was 300.32 µg/mL for the aqueous extract, and 76.60 µg/mL for the ethanolic extract. The 24 h LC50 was 1405.96 µg/mL for the aqueous extract, and it was 356.32 µg/mL for the ethanolic extract. The aqueous extract was nontoxic to A. salina nauplii, and therefore, it can be used as a possible food additive in fish feed. The results also demonstrated that the different solvents used in the extraction affected the yield and the total phenolic, total flavonoid, and condensed tannin content. Further in vivo and cell line cytotoxicity testing is recommended to substantiate these findings.

An Evaluation Research About Effects of Characterized Cadmium Selenide (CdSe) and Lead Selenide (PbSe) Quantum Dots on Brine Shrimp (Artemia salina).

Quantum dots (QDs), such as cadmium selenide (CdSe) and lead selenide (PbSe) exhibit excellent optical, magnetic and chemical properties due to their extremely size (ca. 1-10 nm) and are attractive semiconductor nanomaterials for optical studies and energy storage. In this study, aqueous synthesis of CdSe and PbSe QDs in a size range of 2-10 nm was described. Synthesized QDs were characterized using SEM and TEM, DLS, zeta potential, FTIR, EDX and XRD. Highest accumulation (72.5 ± 5.8 mg L-1) of PbSe QDs occurred at 10 ppm suspensions. In general accumulation increased up to 48 h exposure then fluctuate tended to decline. For CdSe QDs, accumulation tended to decrease for 72 h exposure except that for 5 ppm groups. For the elimination period, in general, the elimination levels of PbSe and CdSe QDs from exposed individuals decreased (p < 0.05) even it has some fluctuate.

Transcriptomic responses of Artemia salina exposed to an environmentally relevant dose of Alexandrium minutum cells or Gonyautoxin2/3.

Toxicities of the marine algae Alexandrium minutum and its excreted gonyautoxins (GTXs) to the marine crustacean Artemia salina were investigated. Mortality was observed for neither larvae nor adult A. salina exposed to A. minutum at a density of 5000 cells/mL or 0.5 µM GTX2/3. After exposure, the full transcriptome of adult A. salina was assembled and functionally annotated. A total of 599,286 transcripts were obtained, which were clustered into 515,196 unigenes. Results of the transcriptional effect level index revealed that direct exposure to the toxic algae A. minutum caused greater alterations in the transcriptome than did exposure to the extracellular product GTX2/3. Mechanisms of effects were different between exposure of A. salina to A. minutum cells or GTX2/3. Exposure to A. minutum modulated formation of the ribonucleoprotein complex and metabolism of amino acids and lipids in A. salina. Exposure to GTX2/3 exposure inhibited expression of genes related to metabolism of chitin, which might result in disruption of molting process or disturbed sheath morphogenesis. Overall, effects on transcription observed in this study represent the first report based on application of next generation sequencing techniques to investigate the transcriptomic response of A. salina exposed to an environmentally realistic level of A. minutum or GTX2/3.

Nanoecotoxicology study of the response of magnetic O-Carboxymethylchitosan loaded silver nanoparticles on Artemia salina.

Magnetic silver nanoparticles (MNPAg) are interesting nanotechnology materials with borderless environmental science, that can be used to disinfect water contaminated with pathogenic bacteria. The use of MNPAg leads to increased risk of nanomaterial contamination in the environment, especially natural water sources, with harmful effects on the ecosystem. This study investigating survival and enzyme activity of magnetic O-carboxymethylchitosan loaded silver nanoparticle on Artemia salina. The results showed that mortality increased with increasing concentrations of MNPAg. O-Carboxymethylchitosan loaded silver nanoparticles were found to be more toxic, with a LC50 of 902.1 mg/L for γ-Fe2O3/Ag without reducing agent. Accumulation of silver on Artemia salina depends on the type of nanoparticle. Accumulation of nanoparticle containing polymers (carboxymethylchitosan/γ-Fe2O3/Ag without reducing agent, carboxymethylchitosan/γ-Fe2O3/Ag reduced with sucrose and carboxymethylchitosan/γ-Fe2O3/Ag reduced with NaBH4) were found to be higher than γ-Fe2O3/Ag reduced with NaBH4, γ-Fe2O3/Ag reduced with sucrose and γ-Fe2O3/Ag without reducing agent under the same experimental conditions. The antioxidant enzyme (CAT, SOD and GST) activities increased slightly following exposure, indicating that the toxic effects are related to oxidative stress. The combined results so far indicate that MNPA does not have the potential to affect aquatic organisms when released into the ecosystem.

Ingestion and bioaccumulation of polystyrene nanoplastics and their effects on the microalgal feeding of Artemia franciscana.

Nanoplastics (NPs) have become one of the most serious environmental problems nowadays. The environmental issues linked to NPs are attributed to the effects after ingestion in marine organisms. Due to the incipient and controversial information about the effects of PS NPs on the feeding of organisms, the aim of this work is to assess (i) digestion dynamics of Artemia franciscana when exposed to PS NPs as the lowest concentration of PS NPs reported in toxicity test [0 (control), 0.006 and 0.6 mg·L-1] and possible interferences in the ingestion of microalgae and (ii) the accumulation and depuration of PS NPs by A. franciscana. Artemia were subjected to ingestion experiments [24 h and 3.5 h], in which the organisms were exposed to PS NPs or to PS NPs + microalgae. Post-exposure feeding (24 h exposure and 2 h feeding) and depuration (24 h exposure and 24 h of depuration) were also carried out. More than 90% of the PS NPs were ingested by Artemia and bioaccumulated in the mandible, stomach, gut, tail gut and appendages after 24 h. The ingestion of microalgae was not affected by the presence of the PS NPs. Data of post-exposure feeding indicated that Artemia previously exposed to plastic and/or microalgae presented similar microalgal ingestion (around 70%); the highest microalgal consumption (around 90%) was recorded in the treatment in which Artemia were previously starved (no plastic and no microalgae). The presence of PS NPs in the gut after the depuration experiments indicates that 24 h was not enough to eliminate the PS NPs.

TDCIPP exposure affects Artemia franciscana growth and osmoregulation.

Environmental monitoring has demonstrated widespread occurrence of the flame-retardant tris(1,3-dichloro-2-propyl) phosphate (TDCIPP), raising concerns about the impact on aquatic life. Using 1H NMR and GC-MS metabolomics and 20-day body length experiments, we have determined that exposure to TDCIPP affects Artemia franciscana. The LC50 for a 48 h TDCIPP exposure was determined to be 37.1 ±â€¯1.3 µM. Acute exposure (48 h) to 20.0 µM did not affect A. franciscana body length but did elicit a metabolic change. Chronic exposure to 0.50 µM TDCIPP caused decreased body length in A. franciscana exposed for 20 days and elicited a metabolic response. Principal component analysis revealed variance between acute and chronic exposure along PC1 (36.4%) and between control and TDCIPP along PC2 (17.4%). One-way ANOVA indicated that 19 metabolites were significantly affected by TDCIPP exposure; namely metabolites of the osmolyte class, including betaine, phosphocholine, gadusol, taurine, glycerol and trehalose - metabolites that are essential osmoprotectants in extremophile species. Other pathways that may be perturbed by TDCIPP exposure include one carbon, glycine, serine, threonine, and glycerophospholipid metabolism.

Time-dependent effects of polystyrene nanoparticles in brine shrimp Artemia franciscana at physiological, biochemical and molecular levels.

Micro- (<5 mm) and nanoplastics (<1 µm) are emerging threats for marine ecosystems worldwide. Brine shrimp Artemia is recognized as a suitable model among planktonic species for studying the impact of polystyrene nanoparticles (PS NPs) through short and long-term bioassays. Our study aims to evaluate the time-dependent effects of cationic amino-modified PS-NH2 (50 nm) in A. franciscana after short- (48 h) and long-term exposure (14 days). For this purpose, nauplii were exposed to a concentration range of PS-NH2 (0.1, 1, 3 and 10 µg/mL) in natural sea water (NSW), and physiological, biochemical and molecular responses were investigated. Short-term exposure to PS-NH2 caused a decrease in nauplii growth and affected the development in a concentration-dependent manner, long-term exposure impaired the survival, but not the growth and feeding behavior. Oxidative stress was detected after short term exposure as the decrease in the activity of antioxidant enzymes, and was fully evident in the long-term as lipid peroxidation, suggesting an accumulative effect. The decrease in Cholinesterase (ChE) activity observed indicates possible neurotoxic action of PS-NH2. Also, Carboxylesterase (CbE) inhibition by PS-NH2, described for the first time in this study, anticipates potential effects in biotransformation of exogenous and endogenous compounds, being the crustacean juvenile hormone methyl farnesoate (MF) that regulates development and molting, one candidate. Furthermore, short- and long-term exposure to PS-NH2 affect the expression of genes involved in cell protection, development and molting. Overall, our results reveal that low PS-NH2 concentrations induce physiological, biochemical and molecular (changes in gene expression) alterations in Artemia, and point at their potential risk for this model organism, supporting the general concern about nanoplastics occurrences in aquatic environments and their ability to represent an ecological threat for aquatic zooplanktonic species.

The function of supplemental foods for improved crop establishment of generalist predators Orius insidiosus and Dicyphus hesperus.

As with many biological control agents, generalist predators rarely survive prolonged periods of prey scarcity. Towards improving crop establishment of two major predators used in North America, Orius insidiosus and Dicyphus hesperus, this study examined the role of supplemental foods in achieving greater predator survival and faster development. In controlled environment trials, developmental time and survival were compared for predators offered diets including Ephestia eggs, Artemia cysts, Typha pollen, or combinations of these. Nymphal developmental time was significantly shorter and survival greater for both predators reared on diets that included Ephestia eggs. Interestingly, D. hesperus could successfully complete nymphal development on Artemia cysts whereas O. insidiosus could not, alluding to fundamental physiological differences between these predators. In greenhouse assays, D. hesperus was more abundant after six weeks when offered diets that included Ephestia eggs either alone or in combination with pollen or Artemia cysts relative to other diets. In contrast, only diets of Ephestia eggs, Typha pollen or their combination could significantly increase O. insidiosus crop abundance relative to the unfed control. Together, this work highlights important differences in the relative values of supplemental foods for generalist predators used in crop protection. It is also meaningful in guiding biocontrol practitioners globally in the rapidly growing sector of greenhouse vegetable production.

Microplastics disturb the anthozoan-algae symbiotic relationship.

World production of plastic has dramatically increased from the 1950's and now it reaches approximately 311 million tons per year. The resulting accumulation of small plastic detritus less than 5 mm in size, termed "microplastics", has started threatening the life cycles of marine organisms. Here we show the first evidence that microplastics disturb the initiation of symbiotic relationships in anthozoan-algae symbiosis. We found in both the aposymbiotic sea-anemone Aiptasia sp. and the coral Favites chinensis that the infectivity of symbiotic algae into the host is severely suppressed by microspheres fed either directly or indirectly through microsphere-fed Artemia sp. Similar trends were seen when microplastics collected from commercial facewash were used instead of microspheres. Therefore, ongoing accumulation of microplastics in the ocean might disturb the healthy anthozoan-algae symbiotic relationships, which are cornerstones of the biologically enriched coral reef ecosystem.